Vaginitis/vaginosis is one of the most common conditions for women to seek medical care. Some 10 million office visits are attributed to vaginitis, which accounts for a health care cost of over $500 million each year. Over 90% of vaginitis can be attributed to three types of vaginitis: bacterial vaginosis (BV), Candida vaginitis (CV) and Trichomoniasis vaginitis (TV). In the past, we have developed a point-of-care (POC) BV test using a Homogeneous Biochemiluminescence Assay (HBA)-based technology. The HBA technology enables a simple (one-step assay), sensitive, and rapid (<10 min.) test that uses a simple and inexpensive hand-held detector. In this application, we propose to develop two additional vaginosis POC tests so it will cover testing majority of the vaginosis cases using this samples tests. In this proposal, we will test whether two prototypes of these HBA-based assays can be used the same way as the BV test for sensitive and specific detection of CV and TV, respective, in POC settings. The proposal contains the following four specific aims: 1. Synthesis and Purification of Two Substrates for CV and TV Detections 2. Assay Optimization and Test Kit Formulation 3. Evaluation of Analytical Performance 4. Clinical Assessment of the Proposed Tests The proposed studies will demonstrate whether the detection technology (the HBA technology) and the two novel markers we adapted in this new development are appropriate for rapid and accurate diagnosis of CV and TV. Successful outcomes from the proposed studies will enable the development of FDA approved CV and TV diagnostic tests that would improve women's health care in U.S. and abroad, particularly in resources poor regions. PUBLIC HEALTH RELEVANCE: Women often seek medical care due to vaginitis/vaginosis, which accounts about 10 million office visits with a health care cost of over $500 million each year. Over 90% of those vaginitis are caused by three specific types of vaginitis, i.e., bacterial vaginosis (BV), Candida vaginitis (CV) and Trichomoniasis vaginitis (TV). If easy-to- use and accurate tests can be developed to allow rapid detections of these 3 conditions, it would save medical care cost and improve women's health as these conditions can be readily treated. So far, we have already developed the BV test. In this application, we propose to develop CV and TV tests using the same technology. Successful outcomes from the proposed studies - evaluation of a novel detection technology and two novel markers for rapid and accurate diagnosis of Candidiasis and Trichomoniasis - will markedly improve the diagnosis of these two common types of vaginitis, which affect hundreds of millions of women each year, thereby improving the women's health care here in U.S. and globally.